Compared to confocal imaging, multi-photon microscopy allows cells to survive longer during the imaging process, due to reduced photobleaching and phototoxicity
Bio-Rad Cell Science Division is set to expand the boundaries of live cell research with the launch of the Radiance2100 MP Rainbow laser scanning imaging system, the first and only multi-photon system to incorporate full spectral imaging capabilities. Compared to confocal imaging, multi-photon microscopy allows cells to survive longer during the imaging process, due to reduced photobleaching and phototoxicity.
The longer wavelength illumination light also penetrates more easily into biological tissue and allows images to be resolved at greater depths. Vitally, multi-photon microscopy produces useful images even if the emitted fluorescence is scattered as it leaves the sample as the optical sectioning and image resolution are determined by the illumination process.
Traditional spectral separation techniques rely upon using a dispersive element that requires a low acceptance numerical aperture to maintain spectral resolution. The Radiance2100 MP Rainbow has been specifically designed to avoid such restrictions by utilising advances in dielectric technology.
With filters it is possible to open the confocal iris to the fully open position and in combination with the signal enhancing lens system (Sels) it is possible to collect significantly more scattered light than a dispersive system, without effecting spectral resolution.
In addition to the filter flexibility, the Rainbow configuration maintains separate gain controls for each channel, allowing the balancing of different intensity fluorophores without damaging the sample.
When collection of pure channels is not possible, Bio-Rad's SpectraSharp software can create separate images through spectral reassignment.
This is especially useful for significantly overlapping fluorescent proteins, unavoidable co-excitation and the overlapping of autofluorescent signals and fluorophore emission.
The SpectraSharp software does not require a spectral series and therefore eliminates the need for time-consuming collections and allows spectral reassignment of images collected using direct detectors.
The complete Radiance2100 MP Rainbow system, plus a range of accessories, is available from Bio-Rad. Existing Radiance2100, Radiance2100 Rainbow and Radiance2100 MP users can upgrade to full multi-photon spectral analysis functionality.
